Ultra-compact fiber-optic two-photon microscope for functional fluorescence imaging in vivo

Opt Express. 2008 Apr 14;16(8):5556-64. doi: 10.1364/oe.16.005556.

Abstract

We present a small, lightweight two-photon fiberscope and demonstrate its suitability for functional imaging in the intact brain. Our device consists of a hollow-core photonic crystal fiber for efficient delivery of near-IR femtosecond laser pulses, a spiral fiber-scanner for resonant beam steering, and a gradient-index lens system for fluorescence excitation, dichroic beam splitting, and signal collection. Fluorescence light is remotely detected using a standard photomultiplier tube. All optical components have 1 mm dimensions and the microscope's headpiece weighs only 0.6 grams. The instrument achieves micrometer resolution at frame rates of typically 25 Hz with a field-of-view of up to 200 microns. We demonstrate functional imaging of calcium signals in Purkinje cell dendrites in the cerebellum of anesthetized rats. The microscope will be easily portable by a rat or mouse and thus should enable functional imaging in freely behaving animals.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cerebellum / ultrastructure*
  • Dendrites / ultrastructure*
  • Fiber Optic Technology / instrumentation*
  • Image Enhancement / instrumentation*
  • Image Enhancement / methods
  • Mice
  • Microscopy, Fluorescence, Multiphoton / instrumentation*
  • Microscopy, Fluorescence, Multiphoton / methods
  • Miniaturization
  • Optical Fibers
  • Rats